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A Schweiger

Publications and source records attributed to A Schweiger.

At least 19 recordsLinked to original sources
PubMed2006

Analyzing heme proteins using EPR techniques: the heme-pocket structure of ferric mouse neuroglobin.

In this work, an electron paramagnetic resonance (EPR) strategy to study the heme-pocket structure of low-spin ferric heme proteins is optimized. Frozen solutions of ferric mouse neuroglobin (mNgb) are analyzed by means of electron spin echo envelope modulation and pulsed electron-nuclear double resonance techniques. The hyperfine and nuclear quadrupole couplings of the directly coordinating heme and histidine nitrogens are derived and are discussed in comparison with known data of other ferric porphyrin compounds. In combination with the hyperfine matrices of the imidazole protons, the 14N EPR parameters reveal structural information on the heme pocket of mNgb that is in agreement with previous X-ray diffraction data on neuroglobins.

Animals↗
PubMed2002

Numerical simulation of one- and two-dimensional ESEEM experiments.

Numerical simulation has become an indispensable tool for the interpretation of pulse EPR experiments. In this work it is shown how automatic orientation selection, grouping of operator factors, and direct selection and elimination of coherences can be used to improve the efficiency of time-domain simulations of one- and two-dimensional electron spin echo envelope modulation (ESEEM) spectra. The program allows for the computation of magnetic interactions of any symmetry and can be used to simulate spin systems with an arbitrary number of nuclei with any spin quantum number. Experimental restrictions due to finite microwave pulse lengths are addressed and the enhancement of forbidden coherences by microwave pulse matching is illustrated. A comparison of simulated and experimental HYSCORE (hyperfine sublevel correlation) spectra of ordered and disordered systems with varying complexity shows good qualitative agreement.

Journal Article↗
PubMed2001

Highly efficient phosphodiester hydrolysis promoted by a dinuclear copper(II) complex.

The interaction of Cu(II) with the ligand tdci (1,3,5-trideoxy-1,3,5-tris(dimethylamino)-cis-inositol) was studied both in the solid state and in solution. The complexes that were formed were also tested for phosphoesterase activity. The pentanuclear complex [Cu(5)(tdciH(-2))(tdci)(2)(OH)(2)(NO(3))(2)](NO(3))(4).6H(2)O consists of two dinuclear units and one trinuclear unit, having two shared copper(II) ions. The metal centers within the pentanuclear structure have three distinct coordination environments. All five copper(II) ions are linked by hydroxo/alkoxo bridges forming a Cu(5)O(6) cage. The Cu-Cu separations of the bridged centers are between 2.916 and 3.782 A, while those of the nonbridged metal ions are 5.455-5.712 A. The solution equilibria in the Cu(II)-tdci system proved to be extremely complicated. Depending on the pH and metal-to-ligand ratio, several differently deprotonated mono-, di-, and trinuclear complexes are formed. Their presence in solution was supported by mass, CW, and pulse EPR spectroscopic study, too. In these complexes, the metal ions are presumed to occupy tridentate [O(ax),N(eq),O(ax)] coordination sites and the O-donors of tdci may serve as bridging units between two metal ions. Additionally, deprotonation of the metal-bound water molecules may occur. The dinuclear Cu(2)LH(-3) species, formed around pH 8.5, provides outstanding rate acceleration for the hydrolysis of the activated phosphodiester bis(4-nitrophenyl)phosphate (BNPP). The second-order rate constant of BNPP hydrolysis promoted by the dinuclear complex (T = 298 K) is 0.95 M(-1) s(-1), which is ca. 47600-fold higher than that of the hydroxide ion catalyzed hydrolysis (k(OH)). Its activity is selective for the phosphodiester, and the hydrolysis was proved to be catalytic. The proposed bifunctional mechanism of the hydrolysis includes double Lewis acid activation and intramolecular nucleophilic catalysis.

Catalysis↗
PubMed2001

A Q-band pulse EPR/ENDOR spectrometer and the implementation of advanced one- and two-dimensional pulse EPR methodology.

A versatile high-power pulse Q-band EPR spectrometer operating at 34.5--35.5 GHz and in a temperature range of 4--300 K is described. The spectrometer allows one to perform one- and two-dimensional multifrequency pulse EPR and pulse ENDOR experiments, as well as continuous wave experiments. It is equipped with two microwave sources and four microwave channels to generate pulse sequences with different amplitudes, phases, and carrier frequencies. A microwave pulse power of up to 100 W is available. Two channels form radiofrequency pulses with adjustable phases for ENDOR experiments. The spectrometer performance is demonstrated by single crystal pulse ENDOR experiments on a copper complex. A HYSCORE experiment demonstrates that the advantages of high-field EPR and correlation spectroscopy can be combined and exploited at Q-band. Furthermore, we illustrate how this combination can be used in cases where the HYSCORE experiment is no longer effective at 35 GHz because of the shallow modulation depth. Even in cases where the echo modulation is virtually absent in the HYSCORE experiment at Q-band, matched microwave pulses allow one to get HYSCORE spectra with a signal-to-noise ratio as good as at X-band. Finally, it is shown that the high microwave power, the short pulses, and the broad resonator bandwidth make the spectrometer well suited to Fourier transform EPR experiments.

Journal Article↗
PubMed2001

Longitudinally detected EPR: improved instrumentation and new pulse schemes.

A new setup for longitudinal detection (LOD) of EPR signals based on a commercial pulse EPR spectrometer equipped with an ENDOR probehead is presented. The design is suited for pulse LOD EPR and amplitude-modulated cw LOD EPR experiments. The sensitivity is substantially increased compared with earlier designs. Two new pulse schemes that take full advantage of the special properties of the setup are invented. In transient-nutation longitudinally detected EPR (TN-LOD EPR), the nutation of magnetization during a microwave pulse is used to measure the EPR signal. In pulse-train excited longitudinally detected EPR (PT-LOD EPR), a train of microwave pulses that periodically inverts the magnetization is applied. First experimental results on radicals and metal complexes at room temperature are presented.

Journal Article↗
PubMed2001

Electron paramagnetic resonance evidence for binding of Cu(2+) to the C-terminal domain of the murine prion protein.

Transmissible spongiform encephalopathies in mammals are believed to be caused by scrapie form of prion protein (PrP(Sc)), an abnormal, oligomeric isoform of the monomeric cellular prion protein (PrP(C)). One of the proposed functions of PrP(C) in vivo is a Cu(II) binding activity. Previous studies revealed that Cu(2+) binds to the unstructured N-terminal PrP(C) segment (residues 23-120) through conserved histidine residues. Here we analyzed the Cu(II) binding properties of full-length murine PrP(C) (mPrP), of its isolated C-terminal domain mPrP(121-231) and of the N-terminal fragment mPrP(58-91) in the range of pH 3-8 with electron paramagnetic resonance spectroscopy. We find that the C-terminal domain, both in its isolated form and in the context of the full-length protein, is capable of interacting with Cu(2+). Three Cu(II) coordination types are observed for the C-terminal domain. The N-terminal segment mPrP(58-91) binds Cu(2+) only at pH values above 5.0, whereas both mPrP(121-231) and mPrP(23-231) already show identical Cu(II) coordination in the pH range 3-5. As the Cu(2+)-binding N-terminal segment 58-91 is not required for prion propagation, our results open the possibility that Cu(2+) ions bound to the C-terminal domain are involved in the replication of prions, and provide the basis for further analytical studies on the specificity of Cu(II) binding by PrP.

Animals↗
PubMed2001

Differences in proliferation of the hematopoietic cell line TF-1 and cytokine production by peripheral blood leukocytes induced by 2 naturally occurring forms of human IL-3.

BACKGROUND: A naturally occurring polymorphism in the coding region of the human IL3 gene leads to a change in amino acid residue 8 from proline to serine. OBJECTIVE: We sought to determine whether the 2 different forms of IL-3 varied in function. These different forms are available as recombinant proteins (recombinant human IL-3/proline 8 [rhIL-3/P8] and recombinant human IL-3/serine 8 [rhIL-3/S8]). METHODS: The erythroleukemic cell line TF-1 was incubated with varying concentrations of rhIL-3/P8 or rhIL-3/S8 to determine the capacity of each type of IL-3 to induce proliferation. Human leukocytes were primed with rhIL-3/P8 or rhIL-3/S8 for up to 24 hours and then stimulated with anti-IgE and assessed for leukotrienes (LTs), IL-4, and TNF-alpha. RESULTS: Proliferation of TF-1 cells was induced by both forms of IL-3 at 48 and 72 hours but to a greater degree by rhIL-3/P8. In contrast, the mean fold increase over control values of LT and IL-4 production was higher after priming the cells with rhIL-3/S8 versus rhIL-3/P8. Additionally, TNF-alpha production was greater (and reached significance only) for rhIL-3/S8. This activity was independent of IgE and thus directly stimulated by IL-3. Studies with basophil-enriched and basophil-depleted cell preparations revealed that LT production was evident only from the former and TNF-alpha only from the latter. CONCLUSION: We conclude that the 2 naturally occurring forms of human IL-3 have similar spectra of activities on cells with IL-3 receptors, but the 2 forms have reversed relative efficacies for promoting proliferation (rhIL-3/P8 > rhIL-3/S8) compared with priming or inducing mediator secretion (rhIL-3/S8 > rhIL-3/P8).

Cell Line↗
PubMed2000

Multiphoton resonances in pulse EPR

Two- and three-photon electron spin echoes of a two-level system are observed using, in addition to the microwave excitation, a linearly polarized radio-frequency field oriented along the static magnetic field B(0). Such multiphoton echoes are detected when the sum of the energies of one microwave and one or two radio-frequency photons are equal to the difference between energies of two spin states. The multiphoton character of the echoes is confirmed by measuring the spin nutation frequency as a function of the angle between the radio-frequency field and B(0), and monitoring the echo amplitude as a function of the radiation field strengths. Floquet theory, usually applied for the description of multiphoton resonances with an odd number of photons, is extended to the case where an even number of photons participate in the transition. Copyright 2000 Academic Press.

Journal Article↗
PubMed2000

One- and two-dimensional pulse electron paramagnetic resonance spectroscopy: concepts and applications.

During the last two decades, the possibilities of pulse electron paramagnetic resonance (EPR) and pulse electron nuclear double resonance (ENDOR) spectroscopy have increased tremendously. While at the beginning of the 1980s pulse-EPR and ENDOR applications were still a rarity, the techniques are now very frequently applied in chemistry, physics, materials science, biology and mineralogy. This is mainly due to the considerable efforts invested in the last few years on instrument development and pulse-sequence design. Pulse-EPR spectrometers are now commercially available, which enables many research groups to use these techniques. In this work, an overview of state-of-the-art pulse EPR and ENDOR spectroscopy is given. The rapid expansion of the field, however, does not allow us to give an exhaustive record of all the pulse methods introduced so far. After a brief and very qualitative description of the basic principles of pulse EPR, we discuss some of the experiments in more detail and illustrate the potential of the methods with a number of selected applications.

Bacteriochlorophylls↗
PubMed2000

Cysteine proteinase cathepsin H in tumours and sera of lung cancer patients: relation to prognosis and cigarette smoking.

In order to evaluate the role of cysteine peptidase cathepsin H (Cath H) in human lung cancer its protein levels were determined in 148 pairs of lung tumour tissue and adjacent non-tumourous lung parenchyma using the enzyme-linked immunosorbent assay technique. Additionally, Cath H levels were determined in sera of 171 patients with malignant tumours, 34 patients with benign lung diseases and 47 healthy controls. The median level of Cath H in tumour tissue was 0.64 times that in the corresponding lung parenchyma. Relating tumour levels with histological type we found higher Cath H levels in small-cell and adenocarcinomas and lower levels in squamous cell carcinoma, large-cell carcinoma and secondary tumours. A significant difference in Cath H level between lung tumour tissue and non-tumourous lung parenchyma was associated with the group of cigarette smokers (156 vs 263 ng mg(-1) protein, P < 0.001). For this group of patients Cath H tumour levels correlated with the survival rate, while for the entire patient population this was not the case. Smokers with high tumour levels of Cath H experienced poor survival. Cath H was significantly higher in sera of patients with malignant and benign lung diseases than in control sera (P < 0.001). The increase was significant for all histological types, being the highest in small-cell and squamous cell carcinomas. Our study reveals that in lung tumours there is different behaviour of Cath H compared with other cysteine peptidases, e.g. cathepsin B and cathepsin L. Variations between tissue and serum levels of Cath H indicate either reduced expression or enhanced secretion of this enzyme in lung tumours.

Carcinoma, Non-Small-Cell Lung↗
PubMed1999

Dead time-dependent line distortions in absolute-value electron spin echo envelope modulation spectra.

It is shown that in pulse EPR experiments, absolute-value spectra can be distorted because of the instrumental dead time to an extent that the interpretation of the spectral features becomes impossible. The mathematical background for the description of the distortions is given, and the far-reaching consequences of the effect are illustrated by model calculations. A cross-term averaging procedure is proposed to get rid of these distortions in two-, three-, and four-pulse electron spin echo envelope modulation experiments. The potential of cross-term averaging is demonstrated in two- and three-pulse experiments on a single crystal and a powder sample.

Electron Spin Resonance Spectroscopy↗
PubMed1998

Nutation-Frequency Correlated EPR Spectroscopy: The PEANUT Experiment

The phase-inverted echo-amplitude detected nutation (PEANUT) experiment for the measurement of transient electron spin nutation frequencies is introduced. In this new pulse sequence, nutation refocused to a rotary echo is detected via the amplitude modulation of a primary electron spin echo. Since detection and excitation of the nutation are fully separated in time, experiments at very high nutation frequencies become feasible. The nutation frequencies which are proportional to the transition moment of an EPR transition can be used to label individual EPR lines by an additional parameter. Using a two-dimensional PEANUT experiment which correlates the nutation frequencies with the resonance fields, the interpretation of complicated field-swept EPR spectra can considerably be simplified. A theoretical description of the experiment is given and the inner working of the approach is described. The predicted features of PEANUT spectra are verified experimentally and examples of applications to both ordered and disordered systems are given. Copyright 1998 Academic Press. Copyright 1998 Academic Press

Journal Article↗
PubMed1998

Sensitivity Enhancement by Matched Microwave Pulses in One- and Two-Dimensional Electron Spin Echo Envelope Modulation Spectroscopy

The concept of microwave pulse matching is applied to three-pulse electron spin echo envelope modulation and sublevel correlation (HYSCORE) spectroscopy. Matched pulses enhance the efficiency of forbidden transfers and may drastically increase the signal intensity of basic frequency and combination frequency transitions in these conventional pulse EPR experiments. The theory of matched pulses is extended to the case of strong and largely isotropic hyperfine interactions, and numerical simulations are presented to gain a deeper insight into the inner working of the matched-pulse approach. It is shown that the enhancement of combination frequencies in matched HYSCORE can be used to determine the relative sign of hyperfine coupling constants as well as the number of equivalent nuclei. The enormous capacity of the approach is demonstrated on ordered and disordered systems. In particular, it is shown that in HYSCORE experiments the signal-to-noise ratio improvement for strongly coupled nitrogens and for proton combination peaks may be considerably larger than one order of magnitude, corresponding to a reduction in measuring time of more than a factor of 100. Copyright 1998 Academic Press.

Journal Article↗
PubMed1998

Two-dimensional nuclear-Zeeman-resolved electron spin echo envelope modulation (NZ-ESEEM) spectroscopy.

A two-dimensional electron spin echo envelope modulation (ESEEM) experiment, called nuclear-Zeeman-resolved ESEEM (NZ-ESEEM), that correlates nuclear transition frequencies with nuclear Zeeman frequencies is introduced. NZ-ESEEM is basically a three-pulse ESEEM experiment complemented by a magnetic-field pulse applied during part of the free evolution period between the second and third microwave pulse. The inner working of the new approach is explained and the instrumentation is discussed. The capacity of the method is illustrated by two examples of applications.

Algorithms↗
PubMed1998

Efficient simulation of ESEEM spectra using gamma

Using the Gamma class library for C++ it is possible to write very efficient time-domain simulation programs for echo-detected pulse EPR experiments, such as three-pulse ESEEM and HYSCORE. By grouping sections of time-invariant propagators, the initial density operator and the detection operator can be re-defined, and the simulation transforms to that of a free induction decay which can be evaluated very efficiently using elements from Gamma. Compared to a straightforward implementation, reductions in computing time of up to a factor of 673 have been found. Copyright 1998 Academic Press.

Journal Article↗